On the measurement of multi-component T2 relaxation in cartilage by MR spectroscopy and imaging

The multicomponent T₂ relaxation in bovine nasal cartilage (BNC) was investigated by nuclear magnetic resonance spectroscopy using the Carr-Purcell-Meiboom-Gill (CPMG) sequence and microscopic magnetic resonance imaging (μMRI) method using a CPMG-SE imaging sequence. All experimental data were analyzed by the non-negative least square (NNLS) procedure. Only one T₂ component was found in BNC by both experimental methods (about 113 and 170 ms before and after being enzymatically digested by trypsin). Several experimental and specimen-related factors were investigated in this study, and it was found that some of them could produce artificial multi-component T₂, including the use of the standard MSME imaging sequence at certain imaging gradients.     

Assessment of interstitial water content of articular cartilage with T1 relaxation

The interstitial water content typically increases in the early degeneration of articular cartilage. Previously, T₂ relaxation has been related to water content, yet it is known to be strongly affected by the collagen orientation. Articular cartilage plugs from the bovine patella, femur and tibia (N=20) were mapped for T₁ and T₂ at 9.4 T to test the ability of T₁ relaxation to reflect cartilage water content. As a reference, water and proteoglycan (PG) contents were determined. Significant (P<.01) linear associations were demonstrated between the relaxation rates and tissue water content (R₁: r=−.81, R₂: r=−.60) and PG content (R₁: r=.75). After adjustment for the tissue water content, partial correlation analysis did not show significant associations between the relaxation rates and tissue PG content. After the effect of PGs was removed, significant (P<.05) linear correlation between the relaxation rates and tissue water content (R₁: r=−.48, R₂: r=−.50) was observed. Thus, the spin-lattice relaxation rate is proposed to provide a biomarker for water content in articular cartilage.     

Transverse relaxation mechanisms in articular cartilage

Relaxation rates in the rotating frame (R1rho) and spin-spin relaxation rates (R2) were measured in articular cartilage at various orientations of cartilage layer to the static magnetic field (B0), at various spin locking field strengths and at two different static magnetic field strengths. It was found that R1rho in the deep radial zone depended on the orientation of specimens in the magnet and decreased with increasing the spin locking field strength. In contrast, R1rho values in the transitional zone were nearly independent of the specimen orientation and the spin locking field strength. Measurements of the same specimens at 2.95 and 7.05 T showed an increase of R1rho and most R2 values with increasing B0. The inverse B0 dependence of some R2 values was probably due to a multicomponent character of the transverse magnetization decay. The experiments revealed that the dominant T1rho and T2 relaxation mechanism at B0 < or = 3 T is a dipolar interaction due to slow anisotropic motion of water molecules in the collagen matrix. On average, the contribution of scalar relaxation due to rapid proton exchange in femoral head cartilage at 2.95 T is about 6% or less of the total R1rho at the spin locking field of 1000 Hz.     

The zonal architecture of human articular cartilage described by T2 relaxation time in the presence of Gd-DTPA2-

The depth-wise variation of T(2) relaxation time is known to reflect the collagen network architecture in cartilage, while the delayed Gadolinium Enhanced MRI of Cartilage (dGEMRIC) technique is sensitive to tissue proteoglycan (PG) concentration. As the cartilage PG content varies along the tissue depth, the depth-dependent accumulation of the contrast agent may affect the inherent T(2) of cartilage in a nonconstant manner. Therefore, T(2) and dGEMRIC are typically measured in separate MRI sessions. In the present in vitro MRI study at 9.4 T, depth-wise T(2) profiles and collagenous zone thicknesses as determined from T(2) maps in the absence and presence of Gd-DTPA(2-) (T(2) and T(2Gd), respectively) were compared in samples of intact human articular cartilage (n=65). These T(2) measures were further correlated with birefringence (BF) of polarized light microscopy (PLM) to quantify the ability of MRI to predict the properties of the collagen fibril network. The reproducibility of the T(2) measurement in the current setup was also studied. Typical tri-laminar collagen network architecture was observed both with and without Gd-DTPA(2-). The inverse of BF (1/BF) correlated significantly with both T(2) and T(2Gd) (r=0.91, slope=0.56 and r=0.90, slope=0.63), respectively. The statistically significant linear correlations between zone thicknesses as determined from T(2) and T(2Gd) were r=0.55 (slope=0.49), r=0.74 (slope=0.71) and r=0.95 (slope=0.94) for superficial, middle and deep tissue zones, respectively. Reproducibility of the T(2) measurement was worst for superficial cartilage. Consistent with PLM, T(2) and T(2Gd) measurements reveal highly similar depth-dependent information on collagen network in intact human cartilage. Thus, dGEMRIC and T(2) measurements in one MRI session are feasible for intact articular cartilage in vitro.     

Maximizing MR signal for 2D UTE slice selection in the presence of rapid transverse relaxation

Ultrashort TE (UTE) sequences allow direct visualization of tissues with very short T2 relaxation times, such as tendons, ligaments, menisci, and cortical bone. In this work, theoretical calculations, simulations, and phantom studies, as well as in vivo imaging were performed to maximize signal-to-noise ratio (SNR) for slice selective RF excitation for 2D UTE sequences. The theoretical calculations and simulations were based on the Bloch equations, which lead to analytic expressions for the optimal RF pulse duration and amplitude to maximize magnetic resonance signal in the presence of rapid transverse relaxation. In steady state, it was found that the maximum signal amplitude was not obtained at the classical Ernst angle, but at an either lower or higher flip angle, depending on whether the RF pulse duration or amplitude was varied, respectively.     

Depth and orientational dependencies of MRI T(2) and T(1ρ) sensitivities towards trypsin degradation and Gd-DTPA(2-) presence in articular cartilage at microscopic resolution

Depth and orientational dependencies of microscopic magnetic resonance imaging (MRI) T(2) and T(1ρ) sensitivities were studied in native and trypsin-degraded articular cartilage before and after being soaked in 1 mM Gd-DTPA(2-) solution. When the cartilage surface was perpendicular to B(0), a typical laminar appearance was visible in T(2)-weighted images but not in T(1ρ)-weighted images, especially when the spin-lock field was high (2 kHz). At the magic angle (55°) orientation, neither T(2)- nor T(1ρ)-weighted image had a laminar appearance. Trypsin degradation caused a depth- and orientational-dependent T(2) increase (4%-64%) and a more uniform T(1ρ) increase at a sufficiently high spin-lock field (55%-81%). The presence of the Gd ions caused both T(2) and T(1ρ) to decrease significantly in the degraded tissue (6%-38% and 44%-49%, respectively) but less notably in the native tissue (5%-10% and 16%-28%, respectively). A quantity Sensitivity was introduced that combined both the percentage change and the absolute change in the relaxation analysis. An MRI experimental protocol based on two T(1ρ) measurements (without and with the presence of the Gd ions) was proposed to be a new imaging marker for cartilage degradation.     

Multi-components of T2 relaxation in ex vivo cartilage and tendon

The multi-components of T₂ relaxation in cartilage and tendon were investigated by microscopic MRI (μMRI) at 13 and 26 μm transverse resolutions. Two imaging protocols were used to quantify T₂ relaxation in the specimens, a 5-point sampling and a 60-point sampling. Both multi-exponential and non-negative-least-square (NNLS) fitting methods were used to analyze the μMRI signal. When the imaging voxel size was 6.76 × 10⁻⁴ mm³ and within the limit of practical signal-to-noise ratio (SNR) in microscopic imaging experiments, we found that (1) canine tendon has multiple T₂ components; (2) bovine nasal cartilage has a single T₂ component; and (3) canine articular cartilage has a single T₂ component. The T₂ profiles from both 5-point and 60-point methods were found to be consistent in articular cartilage. In addition, the depletion of the glycosaminoglycan component in cartilage by the trypsin digestion method was found to result in a 9.81–20.52% increase in T₂ relaxation in articular cartilage, depending upon the angle at which the tissue specimen was oriented in the magnetic field.     

Quantitative MRI using T1ρ and T2 in human osteoarthritic cartilage specimens: correlation with biochemical measurements and histology

Purpose

A direct correlation between T_1ρ, T₂ and quantified proteoglycan and collagen contents in human osteoarthritic cartilage has yet to be documented. We aimed to investigate the orientation effect on T_1ρ and T₂ values in human osteoarthritic cartilage and to quantify the correlation between T_1ρ, T₂ vs. biochemical composition and histology in human osteoarthritic cartilage.

Materials and methods

Thirty-three cartilage specimens were collected from patients who underwent total knee arthroplasty due to severe osteoarthritis and scanned with a 3T MR scanner for T_1ρ and T₂ quantification. Nine specimens were scanned at three different orientations with respect to the B₀: 0°, 90° and 54.7°. Core punches were taken after MRI. Collagen and proteoglycan contents were quantified using biochemical assays. Histology sections were graded using Mankin scores. The correlation between imaging parameters, biochemical contents and histological scores were studied.

Results

Both mean T_1ρ and T₂ at 54.7° were significantly higher than those measured at 90° and 0°, with T_1ρ showing less increase compared to T₂. R_1ρ (1/T_1ρ) values had a significant but moderate correlation with proteoglycan contents (R=.45, P=.002), while R₂ (1/T₂) was not correlated with proteoglycan. No significant correlation was found between relaxation times (T_1ρ or T₂) and collagen contents. The T_1ρ values of specimen sections with high Mankin scores were significantly higher than those with low Mankin scores (P<.05).

Conclusions

Quantitative MRI has a great potential to provide noninvasive imaging biomarkers for cartilage degeneration in osteoarthritis.     

Magnetic resonance T1ρ imaging of osteoarthritis: a rabbit ACL transection model

Objective

The objective of this study was to develop quantitative T_1ρ-weighted magnetic resonance imaging methodology for the detection and characterization of cartilage degeneration in a rabbit anterior cruciate ligament (ACL) transection model.

Methods

The right knee ACLs of 18 adult female New Zealand white rabbits were transected. The left knee joint served as a sham control. The rabbits were euthanized at 3 (Group 1), 6 (Group 2) and 12 (Group 3) weeks postoperatively. High-resolution 3D fat-saturated spoiled gradient echo images and T_1ρ-weighted images were obtained in both the sagittal and axial planes at 3 T using a quadrature wrist coil. Following MR analysis, histological slides from the lateral femoral condyle cartilage were graded using the Mankin grading system.

Results

For all three groups, the average overall T_1ρ values were significantly higher in the ACL-transected knee compared to control knee, and the percentage differences in T_1ρ values between ACL-transected and control increased with the duration of time after transection. The average Mankin score for ACL-transected knees was higher than that for control for each time point, but this difference was statistically significant only for all groups combined.

Conclusions

This study demonstrates the feasibility of using T_1ρ-weighted imaging as a useful tool in the detection and quantification of cartilage damage in all knee compartments in an ACL-transected rabbit model of cartilage degeneration.     

Single-scan longitudinal relaxation measurements in high-resolution NMR spectroscopy

Several single-scan experiments for the measurement of the longitudinal relaxation time (T1) are proposed. These experiments result in fast and accurate determinations of the relaxation rate, are relatively robust to pulse imperfections, and preserve information about the chemical shift. The method used in these experiments is to first encode the T1 values as a spatial variation of the magnetization and then to read out this variation either by applying a weak gradient during acquisition or by sequentially observing different slices of the sample. As a result, it is possible to reduce the time necessary to determine the T1 values by one or two orders of magnitude. This time saving comes at the expense of the signal-to-noise level of the resulting spectrum and some chemical shift resolution.     

Flip-back, an old trick to face highly contrasted relaxation times: application in the characterization of pharmaceutical mixtures by CPMAS NMR

The ¹³C–¹H CPMAS with flip-back pulse NMR experiment is revisited in view of applications to pharmaceutical mixtures. The analysis of the kinetics of relaxation and CP transfer with and without the flip-back pulse shows that a significant gain in ¹³C signal can be expected (thus in experimental time) from the flip-back pulse for protons with long T₁. The gain is of the order of T₁ of the protons expressed in seconds. The experiment is applied on samples with highly contrasted spin-lattice relaxation times T₁ for protons, situation encountered in pharmaceutical mixtures. The application of the flip-back increases significantly the relative signal intensity of the component with the longer T₁, making this component detectable even after using short recycle delays. Therefore, this CPMAS with flip-back experiment could be used routinely to get ¹³C CPMAS NMR spectra of mixtures in constant experimental time and signal-to-noise ratio without the need for optimization of the recycle delays, and for whatever may be the degree of crystallinity of the active principal ingredient (API) and/or excipients.     

Distributions of transverse relaxation times for soft-solids measured in strongly inhomogeneous magnetic fields

The single-sided NMR-MOUSE sensor that operates in highly inhomogeneous magnetic fields is used to record a CPMG ¹H transverse relaxation decay by CPMG echo trains for a series of cross-linked natural rubber samples. Effective transverse relaxation rates 1/T_2,short and 1/T_2,long were determined by a bi-exponential fit. A linear dependence of transverse relaxation rates on cross-link density is observed for medium to large values of cross-link density. As an alternative to multi-exponential fits the possibility to analyze the dynamics of soft polymer network in terms of multi-exponential decays via the inverse Laplace transformation was studied. The transient regime and the effect of the T₁/T₂ ratio in inhomogeneous static and radiofrequency magnetic fields on the CPMG decays were studied numerically using a dedicated C++ program to simulate the temporal and spatial dependence of the CPMG response. A correction factor T₂/T_2,eff is derived as a function of the T₁/T₂ ratio from numerical simulations and compared with earlier results from two different well logging devices. High-resolution T₁–T₂ correlations maps are obtained by two-dimensional Laplace inversion of CPMG detected saturation recovery curves. The T₁–T₂ experimental correlations maps were corrected for the T₁/T₂ effect using the derived T₂/T_2,eff correction factor.     

Two-dimensional correlation relaxation studies of cement pastes

Two-dimensional nuclear magnetic resonance relaxation correlation studies of cement pastes have been performed on a unilateral magnet, the Surface GARField. Through these measurements, the hydration process can be observed by monitoring the evolution of porosity. Characteristic relaxation time distributions have been observed in different cement pastes: fresh white cement, prehydrated white cement and ordinary Portland cement. The observed T(1)/T(2) ratio in these cements has been shown to agree with expectations based on high field values.     

2H T2rho relaxation dynamics and double-quantum filtered NMR studies

In this study 2H T2rho DQF NMR spectra of water in MCM-41 were measured. The T2rho double-quantum filtered (DQF) NMR signal is generated by applying a radio frequency (RF) field for various durations and then observed after a monitor RF pulse. It was found that the transfer between different quantum coherences by the couplings during long-duration RF fields (i.e., soft pulses) and that residual quadrupolar interaction dominates the signal decay. Knowledge of coherence transfer during long-RF pulses has special significance for the development of sophisticated multi-quantum NMR experiments especially multi-quantum MRI applications.     

In vivo multiecho T2 relaxation measurements using variable TR to decrease scan time

Multiecho T2 relaxation measurements to determine geometric mean T2 (GMT2) and myelin water fraction (MWF) are lengthy, resulting in increased motion artefacts from patient discomfort and reduced patient compliance. The goal of this study was to shorten the acquisition time for multiecho T2 measurements without affecting T1 weighting by varying TR across k-space. Six phantoms and 10 healthy volunteers were imaged with both a constant TR and a variable TR multiecho T2 sequence. T1 weighting was determined by TR at the center of k-space; for variable TR measurement, TR was shortened linearly from the center to the edges of k-space. Phantoms showed excellent agreement for proton density and GMT2 between constant and variable TR measurements. No significant differences were found in proton density or MWF for any of the brain structures between the two measurements. The average GMT2 over all structures between the two experiments was not significantly different. In summary, with the variable TR approach, scan time was reduced by >20%, with minimal loss of image resolution and no significant affect on proton density, MWF or GMT2.     

Determination of blood longitudinal relaxation time (T1) at high magnetic field strengths

In this study, a circulation system was used to measure T(1) values of bovine blood under physiological conditions at field strengths of 4.7, 7 and 9.4 T. Results show that T(1) increases linearly with magnetic field B(0) and can be described with the equation T(1)=129 ms/T B(0)+1167 ms for magnetic field strengths between 1.5 and 9.4 T.     

Improvement of duty-cycle heating compensation in NMR spin relaxation experiments

To reliably measure NMR relaxation properties of macromolecules is a prerequisite for precise experiments that identify subtle variations in relaxation rates, as required for the determination of rotational diffusion anisotropy, CSA tensor determination, advanced motional modeling or entropy difference estimations. An underlying problem with current NMR relaxation measurement protocols is maintaining constant sample temperature throughout the execution of the relaxation series especially when rapid data acquisition is required. Here, it is proposed to use a combination of a heating compensation and a proton saturation sequence at the beginning of the NMR relaxation pulse scheme. This simple extension allows reproducible, robust and rapid acquisition of NMR spin relaxation data sets. The method is verified with (15)N spin relaxation measurements for human ubiquitin.     

Novel S-Gal analogs as H MRI reporters for in vivo detection of β-galactosidase

The quantitative assessment of gene expression and related enzyme activity in vivo could be important for the characterization of gene altering diseases and therapy. The development of imaging techniques, based on specific reporter molecules may enable routine non-invasive assessment of enzyme activity and gene expression in vivo. We recently reported the use of commercially available S-Gal^® as a β-galactosidase reporter for ¹H MRI, and the synthesis of several S-Gal^® analogs with enhanced response to β-galactosidase activity. We have now compared these analogs in vitro and have identified the optimal analog, C3-GD, based on strong T₁ and T₂ response to enzyme presence (ΔR₁ and ΔR₂ ~ 1.8 times S-Gal^®). Moreover, application is demonstrated in vivo in human breast tumor xenografts. MRI studies in MCF7-lacZ tumors implanted subcutaneously in athymic nude mice (n = 6), showed significant reduction in T₁ and T₂ values (each ~ 13%) 2 h after intra-tumoral injection of C3-GD, whereas the MCF7 (wild type) tumors showed slight increase. Thus, C3-GD successfully detects β-galactosidase activity in vivo and shows promise as a lacZ gene ¹H MR reporter molecule.     

Effects of intra- and extracellular space properties on diffusion and T(2) relaxation in a tissue model

The purpose of this study was to investigate the effects of biophysical factors on the diffusion and the relaxation time T(2) independently. Certain properties of the extracellular and the intracellular space may change radically in pathological conditions resulting in water diffusion changes. A tissue model consisting of red blood cells was studied. The extra- and intracellular spaces were modified osmotically and by suspending medium concentration. Diffusion measurements were evaluated with regard to the effective medium theory. Neither the nature of the protein in the extracellular space nor an increased level of intracellular hydration caused a significant net water diffusion change in the cell suspension. The relaxation time T(2) exhibited very little dependence on the extracellular volume fraction or the concentration or the nature of the protein in the extracellular space. An increased level of intracellular hydration resulted in systematically larger T(2) values. It seems probable that increases in extracellular protein concentrations or in the extent of intracellular hydration do not play a significant role in the diffusion changes detected in pathological conditions. T(2) appears to depend on the level of hydration or the total water content but is seemingly less dependent of the concentration and the nature of the extracellular protein in our model solutions.     

Oxygen-induced changes in longitudinal relaxation times in skeletal muscle

The objective was to measure the effect of 100% oxygen inhalation on T1 relaxation times in skeletal muscle. Healthy volunteers were scanned using three different MRI protocols while breathing medical air and 100% oxygen. Measurements of T1 were made from regions of interest (ROIs) within various skeletal muscle groups. Dynamic data of subjects breathing a sequence of air-oxygen-air allowed the calculation of characteristic wash-in and -out times for dissolved oxygen in muscle. Contrary to previous findings, a statistically significant decrease in T1 in skeletal muscle was observed due to oxygen inhalation. We report approximate baseline characteristic values for the response of skeletal muscle to oxygen inhalation. This measurement may provide new biomarkers for evaluation of oxygen delivery and consumption in normal and diseased skeletal muscle.